Objective To investigate the role and underlying mechanism of C1q-like protein 4 (C1ql4) in regulating the characteristics of breast cancer stem cells. Methods qRT-PCR was used to detect the expression of C1ql4 in breast cancer and normal breast epithelial cell lines, as well as to verify the transfection efficiency of C1ql4. Western blot analysis was employed to examine the phosphorylation levels of AKT, IKK, and IκB in different groups. An AKT activator was added to MDA-MB-231 cells with C1ql4 knockdown, whereas inhibitors targeting AKT, IKK, IκB, and NF-κB nuclear translocation were separately introduced to C1ql4-overexpressing MCF-7 cells. The nuclear translocation of NF-κB, expression levels of the target genes TNF-α and IL-1β, formation ability of tumorspheres, and proportion of CD44⁺/CD24^−/low stem-like subgroups were analyzed. Results C1ql4 expression in breast cancer cell lines was significantly upregulated compared with that in normal breast epithelial cells. Western blot analysis showed that p-AKT/AKT, p-IKK/IKK, and p-IκB/IκB ratios markedly reduced in C1ql4-knockdown MDA-MB-231 cells (all P<0.05) but significantly increased in C1ql4-overexpressing MCF-7 cells (all P<0.05). Rescue experiments demonstrated that the addition of an AKT activator to C1ql4-knockdown MDA-MB-231 cells resulted in the enhanced nuclear translocation of NF-κB, the increased nuclear/cytoplasmic NF-κB ratios, the elevated TNF-α and IL-1β expression levels, and significant recovery of tumorsphere formation ability and the proportion of CD44⁺/CD24^−/low stem-like subpopulations (all P<0.05). Conversely, in C1ql4-overexpressing MCF-7 cells, treatment with AKT, IKK, IκB, or NF-κB nuclear translocation inhibitors led to a reduction in NF-κB nuclear translocation, decreased nuclear/cytoplasmic NF-κB ratios, and declines in TNF-α and IL-1β expression levels, tumorsphere formation ability, and the CD44⁺/CD24^−/low subpopulation (all P<0.05). Conclusion C1ql4 promotes the translocation of NF-κB from the cytoplasm to the nucleus through the PI3K/AKT/NF-κB signaling pathway and enhances the expression of stemness in breast cancer cells.

BACKGROUND:In recent years,the application of exosomes of periodontal ligament stem cells in periodontal tissue regeneration engineering has been widely studied,but the effect of exosomes on periodontal ligament stem cells derived from inflammatory environment is still unclear.OBJECTIVE:To investigate the effects of exosomes secreted by periodontal ligament stem cells from healthy and inflammatory environments on the proliferation and differentiation of periodontal ligament stem cells from inflammatory environments.METHODS:Human periodontal ligament stem cells from healthy and inflammatory tissues were isolated and cultured by enzyme digestion method.Exosomes were extracted from two kinds of periodontal ligament stem cells using ultracentrifugation.Passage 3 periodontal ligament stem cells derived from inflammatory tissue were selected and cultured in three groups.Cells in the blank group were cultured routinely.The healthy exosome group was added with exosomes secreted by peripheral ligament stem cells derived from healthy tissue.The inflammatory exosome group was added with exosomes secreted by human periodontal ligament stem cells derived from inflammatory tissue.Cell proliferation and cloning were detected.The expression of alkaline phosphatase,the formation of mineralized nodules,and the expression of mRNA and protein of genes related to osteogenesis were detected under osteogenic differentiation.RESULTS AND CONCLUSION:(1) CCK-8 assay and clonal formation test showed that compared with the blank group,two kinds of exosomes could promote the proliferation and colony formation of periodontal ligament stem cells from inflammatory tissue (P<0.05),and the effect of the healthy exosome group was stronger than that of the inflammatory exosome group (P<0.05).(2) Alkaline phosphatase and alizarin red staining showed that compared with the blank group,the two kinds of exosomes could promote the expression of alkaline phosphatase and the formation of mineralized nodules in periodontal ligament stem cells from inflammatory tissue,and the promoting effect of the healthy exosome group was stronger than that of the inflammatory exosome group.RT-PCR and western blot assay showed that compared with the blank group,the two kinds of exosomes could promote the expression of alkaline phosphatase,RUNX2,and type Ⅰ collagen mRNA and protein in periodontal ligament stem cells from inflammatory tissue (P<0.05).The promoting effect of the healthy exosome group was stronger than that of the inflammatory exosome group (P<0.05).(3) The results showed that exosomes secreted by human periodontal ligament stem cells could promote the proliferation and osteogenic differentiation of periodontal ligament stem cells derived from inflammatory environments,and the promoting effect of exosomes secreted by human periodontal ligament stem cells derived from healthy tissues was better than that from human periodontal ligament stem cells derived from inflammatory tissues.

BACKGROUND:In recent years,the application of exosomes of periodontal ligament stem cells in periodontal tissue regeneration engineering has been widely studied,but the effect of exosomes on periodontal ligament stem cells derived from inflammatory environment is still unclear.OBJECTIVE:To investigate the effects of exosomes secreted by periodontal ligament stem cells from healthy and inflammatory environments on the proliferation and differentiation of periodontal ligament stem cells from inflammatory environments.METHODS:Human periodontal ligament stem cells from healthy and inflammatory tissues were isolated and cultured by enzyme digestion method.Exosomes were extracted from two kinds of periodontal ligament stem cells using ultracentrifugation.Passage 3 periodontal ligament stem cells derived from inflammatory tissue were selected and cultured in three groups.Cells in the blank group were cultured routinely.The healthy exosome group was added with exosomes secreted by peripheral ligament stem cells derived from healthy tissue.The inflammatory exosome group was added with exosomes secreted by human periodontal ligament stem cells derived from inflammatory tissue.Cell proliferation and cloning were detected.The expression of alkaline phosphatase,the formation of mineralized nodules,and the expression of mRNA and protein of genes related to osteogenesis were detected under osteogenic differentiation.RESULTS AND CONCLUSION:(1) CCK-8 assay and clonal formation test showed that compared with the blank group,two kinds of exosomes could promote the proliferation and colony formation of periodontal ligament stem cells from inflammatory tissue (P<0.05),and the effect of the healthy exosome group was stronger than that of the inflammatory exosome group (P<0.05).(2) Alkaline phosphatase and alizarin red staining showed that compared with the blank group,the two kinds of exosomes could promote the expression of alkaline phosphatase and the formation of mineralized nodules in periodontal ligament stem cells from inflammatory tissue,and the promoting effect of the healthy exosome group was stronger than that of the inflammatory exosome group.RT-PCR and western blot assay showed that compared with the blank group,the two kinds of exosomes could promote the expression of alkaline phosphatase,RUNX2,and type Ⅰ collagen mRNA and protein in periodontal ligament stem cells from inflammatory tissue (P<0.05).The promoting effect of the healthy exosome group was stronger than that of the inflammatory exosome group (P<0.05).(3) The results showed that exosomes secreted by human periodontal ligament stem cells could promote the proliferation and osteogenic differentiation of periodontal ligament stem cells derived from inflammatory environments,and the promoting effect of exosomes secreted by human periodontal ligament stem cells derived from healthy tissues was better than that from human periodontal ligament stem cells derived from inflammatory tissues.

Literature related to children's adenoid hypertrophy was retrieved to form an expert questionnaire.According to the group standard writing rules of the China Association of Chinese Medicine,the peer consultation,quality evaluation and suitability eval-uation were completed through three rounds of Delphi expert questionnaire surveys and expert discussion meetings,and the Clinical Practice Guidelines for TCM in Children with Adenoidal Hypertrophy was finally formed.The guidelines have been formulated to clarify the scope of application of the guidelines,normative reference documents,terms and definitions,diagnosis,syndrome differentiation,treatment,prevention and care,and to provide an important reference for the clinical practice and diagnosis and treatment norms of tra-ditional Chinese medicine for children with adenoid hypertrophy.

Objective ToinvestigatethecorrelationbetweenCTimagingfindingsoflungadenocarcinomaandepidermalgrowth factorreceptor(EGFR)genemutation.Methods Theclinicaldataof150lungadenocarcinomapatientsinthehospitalfrom October 2015toOctober2017werecollectedretrospectively.AccordingtotheEGFRgenemutation,thepatientsweredividedintononeffectivemutation group (n=78)andeffective mutationgroup (n=72).Univariateanalysisand multivariate L o g istic regression modelwereperformed toexplorethepredictionsignsofeffectiveEGFRgenemutationinlungadenocarcinoma.Results Univariateanalysisshowedthatthe proportionsoffemalepatients,smokinghistory,CTfindingsofspiculesign,necroticsign,pleuralindentationandnonfibrosisin theeffectivemutationgroupweresignificantlyhigherthanthoseinnoneffectivemutationgroup(P<0.05).However,therewereno significantdifferencesbetweenthesetwogroupsinage,diameteroflesions,locationoflesions,densityoflesions,lobulatedsign, cavitation sign ,air bronchogram and pleuralthickening sign (P>0 .05 ).M ultivariate L o g istic regression analysis showed thatfemale (OR=2.612),spiculesign(OR=2.476),necroticsign(OR=2.846),pleuralindentation(OR=2.221)andnonfibrosis(OR=2.476)were independentpredictorsofeffectiveEGFRgenemutationinlungadenocarcinoma(P<0.05).Conclusion FemaleandlungadenocarcinomaCT findingsofspiculesign,necroticsign,pleuralindentationandnonfibrosisarerelatedtoEGFRgenemutation,whichisofgreatsignificanceto distinguishingwildtypefrom mutanttypeofEGFRgeneandguidingtheclinicaltreatment.

Objective: To investigate the expression of lncRNA01296 in esophageal cancer (EC) tissues and its effect on the proliferation and migration of EC TE-2 cells. Methods:Atotal of 36 pairs of esophageal cancer tissues and corresponding para-cancerous tissues were collected from EC patients admitted to the Department of Thoracic Surgery,Affiliated Hospital of Chengde Medical College from January 2017 to September 2018. The human normal esophageal epithelial (HEEC) cells and human esophageal cancer cell lines ECA109, TE-1 and TE-2 were cultured. qPCR was used to detect the mRNAexpressions of lincRNA01296, SNRPA(small nuclear ribonucleoproteinA) and NGF (nerve growth factor) in EC tissues and cells. Recombinant lentiviral interference vectoror control vector were used to transfect EC cell lines, as sh-lncRNA01296#1,#2 and Mock groups. WB was used to detect the protein expressions of SNRPAand NGF in transfected cells. MTS assay was used to detect cell proliferation, and Transwell assays were used to detect cell invasion and migration of TE-2 cells after transfection. Results: The mRNAexpressions of lncRNA01296, SNRPAand NGF were significantly increased in esophageal cancer tissues and cell lines (all P<0.01), and these expressions in poorly differentiated TE-2 cells were higher than those in highly differentiated ECA109 and TE-1 cells (all P<0.05). The mRNAexpressions of lncRNA01296 and NGF in sh-lncRNA01296#1 and sh-lncRNA01296#2 groups were significantly lower than those in Mock group (all P<0.01), while the mRNAexpression of SNRPAshowed no statistical difference among three groups (P>0.05). The protein expressions of lncRNA01296 and NGF in sh-lncRNA01296#1 and sh-lncRNA01296#2 groups were significantly lower than those in Mock group (all P<0.01). The relative proliferation ability of cells in sh-lncRNA01296#1 and shlncRNA01296#2 groups was significantly lower than that of Mock group at 48 and 72 h after transfection (P<0.05 or P<0.01). The number of invasive cells was (72.0±6.3), (36.6±4.3) and (33.9±3.7) in Mock, sh-lncRNA01296#1 and sh-lncRNA01296#2 groups, respectively; and the number of migrated cells was (85.2±9.9), (47.5±8.1) and (43.8±6.5), respectively, indicating that the numbers of invasive and migrated cells in sh-lncRNA01296#1 and sh-lncRNA01296#2 groups were significantly less than those in Mock group(all P<0.01). Conclusion: lncRNA01296 can up-regulate SNRPAexpression to promote NGF-mediated proliferation and metastasis of EC cells, which may provide new target for the diagnosis and treatment of esophagealcancer.

Objective To examine the characteristics of spontaneous activity of brain regions in bipolar Ⅱ depression patients by using resting state fMRI technology, and explore the pathogenesis of bipolarⅡdepression.Methods In total 21 bipolarⅡdepression patients(BD group)and 21 gender-,age-and education-matched healthy controls(HC group)were recruited.The patients were diagnosed according to the DSM-Ⅳ criteria, and the symptoms were evaluated by the Montgomery-Asberg Depression Rating Scale (MADRS), HAMD17and HAMA. The resting-state data was collected via a 3.0 T GE MRI, and was analyzed using an amplitude of low-frequency fluctuation(ALFF)approach,and two-samples t test was used to analyze the difference between BD group and HC group.Correlation analysis was used between the mean value of ALFF in brain regions which were significantly different from the other regions and the depression severity. The brain areas that were significantly different from the other brain regions were regarded as regions of interest,and were made functional connectivity with the whole brain.The functional connectivity was compared between the two groups to explore the impaired emotional network in bipolar Ⅱdepression. Results The ALFFs of left paracentral lobule(t=3.80),right precuneus(t=3.54)and right precentral gyrus(t=3.66) in BD group were significantly higher than HC group(all P<0.05, GRF correction). In BD group, the ALFFs of the left orbital frontal gyrus (r=0.75) and right orbital frontal gyrus (r=0.80) were positively correlated with the HAMA score.The ALFFs of bilateral gyri rectus(r=0.73)and brain regions near the left insular lobe (r=0.74) were positively correlated with the HAMD17score. The ALFF of region 2 of right cerebellar peduncles (r=-0.65) was negatively correlated with the MADRS score(all P<0.05). The functional connectivity between paracentral lobule, precuneus, precentral gyrus and left middle frontal gyrus, left anterior cingulate gyrus together with its lateral gyrus in BD group was significantly increased relative to HC group (t=4.47,4.07; both P<0.05), while the functional connectivity between the right rectangular shape crack and the cortex around was decreased in BD group (t=4.54,P<0.05). Conclusion In bipolar Ⅱ depression patients, the brain function of left paracentral lobule, right precuneus and right precentral gyrus may be impaired and correlated with severity of depressive.

Objective To examine the characteristics of spontaneous activity of brain regions in bipolar Ⅱ depression patients by using resting state fMRI technology, and explore the pathogenesis of bipolarⅡdepression.Methods In total 21 bipolarⅡdepression patients(BD group)and 21 gender-,age-and education-matched healthy controls(HC group)were recruited.The patients were diagnosed according to the DSM-Ⅳ criteria, and the symptoms were evaluated by the Montgomery-Asberg Depression Rating Scale (MADRS), HAMD17and HAMA. The resting-state data was collected via a 3.0 T GE MRI, and was analyzed using an amplitude of low-frequency fluctuation(ALFF)approach,and two-samples t test was used to analyze the difference between BD group and HC group.Correlation analysis was used between the mean value of ALFF in brain regions which were significantly different from the other regions and the depression severity. The brain areas that were significantly different from the other brain regions were regarded as regions of interest,and were made functional connectivity with the whole brain.The functional connectivity was compared between the two groups to explore the impaired emotional network in bipolar Ⅱdepression. Results The ALFFs of left paracentral lobule(t=3.80),right precuneus(t=3.54)and right precentral gyrus(t=3.66) in BD group were significantly higher than HC group(all P<0.05, GRF correction). In BD group, the ALFFs of the left orbital frontal gyrus (r=0.75) and right orbital frontal gyrus (r=0.80) were positively correlated with the HAMA score.The ALFFs of bilateral gyri rectus(r=0.73)and brain regions near the left insular lobe (r=0.74) were positively correlated with the HAMD17score. The ALFF of region 2 of right cerebellar peduncles (r=-0.65) was negatively correlated with the MADRS score(all P<0.05). The functional connectivity between paracentral lobule, precuneus, precentral gyrus and left middle frontal gyrus, left anterior cingulate gyrus together with its lateral gyrus in BD group was significantly increased relative to HC group (t=4.47,4.07; both P<0.05), while the functional connectivity between the right rectangular shape crack and the cortex around was decreased in BD group (t=4.54,P<0.05). Conclusion In bipolar Ⅱ depression patients, the brain function of left paracentral lobule, right precuneus and right precentral gyrus may be impaired and correlated with severity of depressive.

Objective To investigate the correlation between histone deacetylase 1(HDAC1) and vascular endothelial growth factor(VEGF) in the patients with lung adenocarcinoma.Methods Eighty cases of lung adenocarcinoma in our hospital from August 2014 to April 2016 served as the research subjects,and contemporaneous 80 individuals undergoing healthy physical examination were taken as the control group.The fasting venous blood sample was collected in all subjects.Then serum HDAC1 and VEGF levels were detected by ELISA.The differences of serum HDAC1 and VEGF expression levels were compared between the two groups.The HDAC1 and VEGF expression levels in the patients with different characteristics of lung adenocarcinoma and the relation between serum HDAC1 and VEGF concentrations were analyzed.Furthermore the possible influence factors of HDAC1 protein expression level in the patients with lung adenocarcinoma were analyzed.Results The HDAC1 levels in the control group and observation group were(329.56 ± 23.83) ng/L and(568.20 ± 35.40) ng/L,the difference was statistically significant(t=23.576,P=0.000).The VEGF levels in the control group and observation group were(40.26±9.82)ng/L and(296.56±19.80)ng/L respec tively,the difference was statistically significant(t=31.154,P=0.000).The HDAC1 protein level had statistical difference among different genders,ages,clinical stages and smoking history,the HDAC1 protein level in male,age ＞60 years old,clinical stage Ⅲ,V and patients with smoking history were higher(P＜0.05).The Pearson correlation analysis results showed that serum HDAC1 in the patients with lung adenocarcinoma was positively correlated with VEGF protein concentration(r=0.526,P =0.000).The Logistic regression analysis showed that influence factor of HDAC1 protein expression level in the patients with lung adenocarcinoma was clinical stage.Conclusion The high expression of HDAC1 protein in lung adenocarcinoma patients may also simultaneously regulate the VEGF expression,thus promotes the development of lung adenocarcinoma.